Gade Terence P F, Tucker Elizabeth, Nakazawa Michael S, Hunt Stephen J, Wong Waihay, Krock Bryan, Weber Charles N, Nadolski Gregory J, Clark Timothy W I, Soulen Michael C, Furth Emma E, Winkler Jeffrey D, Amaravadi Ravi K, Simon M Celeste
From the Penn Image-Guided Interventions Laboratory (T.P.F.G., S.J.H., C.N.W., G.J.N.), Department of Radiology (T.P.F.G., S.J.H., C.N.W., G.J.N., T.W.I.C., M.C. Soulen), and Department of Pathology (E.E.F.), Hospital of the University of Pennsylvania, Philadelphia, Pa; Abramson Family Cancer Research Institute, Perelman School of Medicine at the University of Pennsylvania, 421 Curie Blvd, 456 BRB II/III, Philadelphia, PA 19104 (E.T., M.S.N., W.W., B.K., M.C. Simon); Abramson Family Cancer Center (B.K., R.K.A.) and Department of Chemistry (J.D.W.), University of Pennsylvania, Philadelphia, Pa.
Radiology. 2017 Jun;283(3):702-710. doi: 10.1148/radiol.2017160728. Epub 2017 Mar 2.
Purpose To characterize hepatocellular carcinoma (HCC) cells surviving ischemia with respect to cell cycle kinetics, chemosensitivity, and molecular dependencies that may be exploited to potentiate treatment with transarterial embolization (TAE). Materials and Methods Animal studies were performed according to institutionally approved protocols. The growth kinetics of HCC cells were studied in standard and ischemic conditions. Viability and cell cycle kinetics were measured by using flow cytometry. Cytotoxicity profiling was performed by using a colorimetric cell proliferation assay. Analyses of the Cancer Genome Atlas HCC RNA-sequencing data were performed by using Ingenuity Pathway Analysis software. Activation of molecular mediators of autophagy was measured with Western blot analysis and fluorescence microscopy. In vivo TAE was performed in a rat model of HCC with (n = 5) and without (n = 5) the autophagy inhibitor Lys05. Statistical analyses were performed by using GraphPad software. Results HCC cells survived ischemia with an up to 43% increase in the fraction of quiescent cells as compared with cells grown in standard conditions (P < .004). Neither doxorubicin nor mitomycin C potentiated the cytotoxic effects of ischemia. Gene-set analysis revealed an increase in mRNA expression of the mediators of autophagy (eg, CDKN2A, PPP2R2C, and TRAF2) in HCC as compared with normal liver. Cells surviving ischemia were autophagy dependent. Combination therapy coupling autophagy inhibition and TAE in a rat model of HCC resulted in a 21% increase in tumor necrosis compared with TAE alone (P = .044). Conclusion Ischemia induces quiescence in surviving HCC cells, resulting in a dependence on autophagy, providing a potential therapeutic target for combination therapy with TAE. RSNA, 2017 Online supplemental material is available for this article.
目的 对在缺血状态下存活的肝细胞癌（HCC）细胞的细胞周期动力学、化学敏感性以及可能用于增强经动脉栓塞（TAE）治疗效果的分子依赖性进行特征描述。材料与方法 动物研究按照机构批准的方案进行。在标准条件和缺血条件下研究HCC细胞的生长动力学。使用流式细胞术测量细胞活力和细胞周期动力学。通过比色细胞增殖测定法进行细胞毒性分析。使用Ingenuity Pathway Analysis软件对癌症基因组图谱HCC RNA测序数据进行分析。通过蛋白质印迹分析和荧光显微镜测量自噬分子介质的激活情况。在有（n = 5）和无（n = 5）自噬抑制剂Lys05的HCC大鼠模型中进行体内TAE。使用GraphPad软件进行统计分析。结果 与在标准条件下生长的细胞相比，HCC细胞在缺血状态下存活，静止细胞比例增加高达43%（P <.004）。阿霉素和丝裂霉素C均未增强缺血的细胞毒性作用。基因集分析显示，与正常肝脏相比，HCC中自噬介质（如CDKN2A、PPP2R2C和TRAF2）的mRNA表达增加。在缺血状态下存活的细胞依赖自噬。在HCC大鼠模型中，自噬抑制与TAE联合治疗导致肿瘤坏死比单独TAE增加21%（P =.
